Discovery of Novel Non-Cyclam Polynitrogenated CXCR4 Coreceptor Inhibitors

ABSTRACT The infection of CD4-negative cells by variants of tissue culture-adapted human immunodeficiency virus type 1 (HIV-1) or HIV-2 strains has been shown to be mediated by the CXCR4 coreceptor. Here we show that two in vitro-established CD4−/CCR5−/CXCR4+ human pre-T-cell lines (A3 and A5) can be productively infected by wild-type laboratory-adapted T-cell-tropic HIV-1 and HIV-2 strains in a CD4-independent, CXCR4-dependent fashion. Despite the absence of CCR5 expression, A3 and A5 cells were susceptible to infection by the simian immunodeficiency viruses SIVmac239 and SIVmac316. Thus, at least in A3 and A5 cells, one or more of the chemokine receptors can efficiently support the entry of HIV and SIV isolates in the absence of CD4. These findings suggest that to infect cells of different compartments, HIV and SIV could have evolved in vivo to bypass CD4 and to interact directly with an alternative receptor.

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PO 8411 MOLECULAR CHARACTERISATION OF GP41 AND GP120 V3 LOOP IN HIV-1C PATIENTS FAILING SALVAGE THERAPY IN BOTSWANA

BMJ Global Health ◽

10.1136/bmjgh-2019-edc.87 ◽

2019 ◽

Vol 4 (Suppl 3) ◽

pp. A34.1-A34

Author(s):

Nokuthula S Ndlovu ◽

Kaelo Seatla

Keyword(s):

Viral Entry ◽

Salvage Therapy ◽

False Positive Rate ◽

V3 Loop ◽

Coreceptor Usage ◽

Drug Resistant ◽

Resistance Mutations ◽

Global Challenge ◽

Cxcr4 Coreceptor ◽

Hiv 1

BackgroundTriple class drug-resistant HIV-1 infection remains a global challenge in individuals with extensive antiretroviral treatment (ART) experience, in terms of high mortality and probability of onward transmission. New therapeutic options within old and new drug classes are therefore essential. We determined if patients failing salvage therapy in Botswana are eligible for maraviroc (MVC) and enfuvirtide (T20) viral entry inhibitors based on the coreceptor usage and drug-resistant mutations in envelope gp120 and gp41.MethodsA total of 38 deep salvage patients were included in the analysis. We amplified and sequenced gp41 and V3 regions of HIV-1 envelope. Drug resistance mutations were analysed according to the IAS-USA 2017 reference mutation lists. Coreceptor usage was determined using PSSM and Geno2Pheno using a false-positive rate (FPR) of 10%.ResultsAmong 38 participants, 34 (89%) were successfully sequenced and amplified gp41 and 26 (68%) gp120 V3 loop sequences were obtained. Major T20 mutation G36S was obtained in 1/34 samples (5.8%) within the study population. Polymorphisms I169V(97%), I135L(100%), E151A(70.6%) and N42S(70.6%) were detected in HR1 and HR2 of gp41. CXCR4 coreceptor associated use, mutation L34M in gp41 HR1 was detected in 2 samples (5%). Analysis of coreceptor usage showed (17/26) 65.4% use of CCR5, and a (9/26) 34.6% use of the CXCR4 coreceptor.ConclusionA moderately high proportion of treatment-experienced (deep salvage) participants had CXCR4 coreceptor using strains. The use of maraviroc in Botswana would require coreceptor tropism testing. Non-T20 treatment experience in Botswana reduces the prevalence of the major mutations that confer resistance to the drug. T20 is therefore a potential alternative drug for patients failing salvage therapy in Botswana.

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A Cell Line-Based Neutralization Assay for Primary Human Immunodeficiency Virus Type 1 Isolates That Use either the CCR5 or the CXCR4 Coreceptor

Journal of Virology ◽

10.1128/jvi.73.11.8966-8974.1999 ◽

1999 ◽

Vol 73 (11) ◽

pp. 8966-8974 ◽

Cited By ~ 128

Author(s):

Alexandra Trkola ◽

Jamie Matthews ◽

Cynthia Gordon ◽

Tom Ketas ◽

John P. Moore

Keyword(s):

Human Immunodeficiency Virus ◽

Cell Line ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Neutralization Assay ◽

Immunodeficiency Virus ◽

A Cell ◽

Cxcr4 Coreceptor ◽

Hiv 1

ABSTRACT We describe here a cell line-based assay for the evaluation of human immunodeficiency virus type 1 (HIV-1) neutralization. The assay is based on CEM.NKR cells, transfected to express the HIV-1 coreceptor CCR5 to supplement the endogenous expression of CD4 and the CXCR4 coreceptor. The resulting CEM.NKR-CCR5 cells efficiently replicate primary HIV-1 isolates of both R5 and X4 phenotypes. A comparison of the CEM.NKR-CCR5 cells with mitogen-activated peripheral blood mononuclear cells (PBMC) in neutralization assays with sera from HIV-1-infected individuals or specific anti-HIV-1 monoclonal antibodies shows that the sensitivity of HIV-1 neutralization is similar in the two cell types. The CEM.NKR-CCR5 cell assay, however, is more convenient to perform and eliminates the donor-to-donor variation in HIV-1 replication efficiency, which is one of the principal drawbacks of the PBMC-based neutralization assay. We suggest that this new assay is suitable for the general measurement of HIV-1 neutralization by antibodies.

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N-Linked Glycosylation of the HIV Type-1 gp120 Envelope Glycoprotein as a Major Determinant of CCR5 and CXCR4 Coreceptor Utilization

Journal of Biological Chemistry ◽

10.1074/jbc.m009779200 ◽

2001 ◽

Vol 276 (16) ◽

pp. 13433-13441 ◽

Cited By ~ 166

Author(s):

Georgios Pollakis ◽

Stanley Kang ◽

Aletta Kliphuis ◽

Moustapha I. M. Chalaby ◽

Jaap Goudsmit ◽

...

Keyword(s):

Envelope Glycoprotein ◽

Major Determinant ◽

Gp120 Envelope ◽

Cxcr4 Coreceptor ◽

Hiv Type 1

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Different Mutational Pathways to CXCR4 Coreceptor Switch of CCR5-Using Simian-Human Immunodeficiency Virus

Journal of Virology ◽

10.1128/jvi.00145-08 ◽

2008 ◽

Vol 82 (11) ◽

pp. 5653-5656 ◽

Cited By ~ 15

Author(s):

Siu-hong Ho ◽

Nataliya Trunova ◽

Agegnehu Gettie ◽

James Blanchard ◽

Cecilia Cheng-Mayer

Keyword(s):

Human Immunodeficiency Virus ◽

Lymph Node ◽

Experimental System ◽

V3 Loop ◽

Infected Macaque ◽

Immunodeficiency Virus ◽

Coreceptor Switch ◽

Cxcr4 Coreceptor ◽

Functional Analyses

ABSTRACT We report here a second case of coreceptor switch in R5 simian-human immunodeficiency virus SF162P3N (SHIVSF162P3N)-infected macaque CA28, supporting the use of this experimental system to examine factors that drive the change in coreceptor preference in vivo. Virus recovered from CA28 plasma (SHIVCA28NP) used both CCR5 and CXCR4 for entry, but the virus recovered from lymph node (SHIVCA28NL) used CXCR4 almost exclusively. Sequence and functional analyses showed that mutations in the V3 loop that conferred CXCR4 usage in macaque CA28 differed from those described in the previously reported case, demonstrating divergent mutational pathways for change in the coreceptor preference of the R5 SHIVSF162P3N isolate in vivo.

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Effects of Human Immunodeficiency Virus Type 1 Infection on CCR5 and CXCR4 Coreceptor Expression on CD4 T Lymphocyte Subsets in Infants and Adolescents

AIDS Research and Human Retroviruses ◽

10.1089/088922204322996545 ◽

2004 ◽

Vol 20 (3) ◽

pp. 305-313 ◽

Cited By ~ 13

Author(s):

Daniel L. Tuttle ◽

Carter R. Coberley ◽

Xiaoming Xie ◽

Zhong C. Kou ◽

John W. Sleasman ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

T Lymphocyte Subsets ◽

Immunodeficiency Virus ◽

Cxcr4 Coreceptor

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CD4-Induced T-20 Binding to Human Immunodeficiency Virus Type 1 gp120 Blocks Interaction with the CXCR4 Coreceptor

Journal of Virology ◽

10.1128/jvi.78.10.5448-5457.2004 ◽

2004 ◽

Vol 78 (10) ◽

pp. 5448-5457 ◽

Cited By ~ 41

Author(s):

Wen Yuan ◽

Stewart Craig ◽

Zhihai Si ◽

Michael Farzan ◽

Joseph Sodroski

Keyword(s):

Human Immunodeficiency Virus ◽

Membrane Fusion ◽

Heptad Repeat ◽

V3 Loop ◽

Repeat Region ◽

Coreceptor Binding ◽

Immunodeficiency Virus ◽

Cxcr4 Coreceptor ◽

Hiv 1

ABSTRACT The synthetic peptide T-20, which corresponds to a sequence within the C-terminal heptad repeat region (HR2) of the human immunodeficiency virus type 1 (HIV-1) gp41 envelope glycoprotein, potently inhibits viral membrane fusion and entry. Although T-20 is thought to bind the N-terminal heptad repeat region (HR1) of gp41 and interfere with gp41 conformational changes required for membrane fusion, coreceptor specificity determined by the V3 loop of gp120 strongly influences the sensitivity of HIV-1 variants to T-20. Here, we show that T-20 binds to the gp120 glycoproteins of HIV-1 isolates that utilize CXCR4 as a coreceptor in a manner determined by the sequences of the gp120 V3 loop. T-20 binding to gp120 was enhanced in the presence of soluble CD4. Analysis of T-20 binding to gp120 mutants with variable loop deletions and the reciprocal competition of T-20 and particular anti-gp120 antibodies suggested that T-20 interacts with a gp120 region near the base of the V3 loop. Consistent with the involvement of this region in coreceptor binding, T-20 was able to block the interaction of gp120-CD4 complexes with the CXCR4 coreceptor. These results help to explain the increased sensitivity of CXCR4-specific HIV-1 isolates to the T-20 peptide. Interactions between the gp41 HR2 region and coreceptor-binding regions of gp120 may also play a role in the function of the HIV-1 envelope glycoproteins.

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Durable Control of HIV-1 Using a Staphylococcus aureus Cas9-Expressing Lentivirus Co-Targeting Viral Latency and Host Susceptibility

10.21203/rs.3.rs-45582/v1 ◽

2020 ◽

Author(s):

Leonard Chavez ◽

Nishith Reddy ◽

Kyle Raymond ◽

Mohamed Bouzidi ◽

Shivani Desai ◽

...

Keyword(s):

Gene Editing ◽

Lentiviral Vector ◽

Viral Reactivation ◽

Host Susceptibility ◽

Target Cells ◽

Gene Therapies ◽

Dual Targeting ◽

Cxcr4 Coreceptor ◽

Infected Cells ◽

Hiv 1

Abstract CRISPR/Cas9 gene editing has the potential to revolutionize the clinical management of HIV-1 infection, and may eliminate the need for antiretroviral therapy (ART). Current gene therapies attempt to either excise HIV-1 provirus or target HIV-1 entry receptors to prevent infection of new cells. Using a viral dynamic model, we determined that combining these two interventions, in the presence or absence of ART, significantly lowers the gene editing efficacy thresholds required to achieve an HIV-1 cure. To implement this dual-targeting approach, we engineered a single lentiviral vector that simultaneously targets multiple highly-conserved regions of the provirus and the host CXCR4 coreceptor, and developed a novel coculture system enabling real-time monitoring of latent infection, viral reactivation, and infection of new target cells. Simultaneous dual-targeting depleted HIV-1-infected cells with significantly greater potency than vectors targeting either virus or host independently, highlighting its potential as an HIV-1 cure strategy.

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